Anti-Myelin PLP antibody (ab28486)

Rabbit polyclonal Myelin PLP antibody. Validated in WB, ELISA, IHC, ICC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Rabbit, Human. Cited in 25 publication(s). Independently reviewed in 12 review(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to Myelin PLP
  • Host species
    Rabbit
  • Specificity
    ab28486 recognises PLP
  • Tested applications
    Suitable for: ELISA, WB, ICC, IHC-FoFr, ICC/IF, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
    Predicted to work with: Cow, Dog, Pig, Macaque monkey
  • Immunogen

    Synthetic peptide:

    CG KGLSATVTGG QKGRGSRG

    , corresponding to amino acids 109-128 of Mouse Myelin PLP

  • Positive control
    • Mouse brain

Properties

Applications

Our Abpromise guarantee covers the use of ab28486 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA 1/10000.
WB 1/1000. Predicted molecular weight: 26,30 kDa.
ICC 1/1000.
IHC-FoFr 1/100.
ICC/IF 1/100.
IHC-P Use at an assay dependent concentration.
IHC-Fr 1/500.
Flow Cyt Use at an assay dependent concentration. PubMed: 19659692

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    This is the major myelin protein from the central nervous system. It plays an important role in the formation or maintenance of the multilamellar structure of myelin.
  • Involvement in disease
    Defects in PLP1 are the cause of leukodystrophy hypomyelinating type 1 (HLD1) [MIM:312080]; also known as Pelizaeus-Merzbacher disease. HLD1 is an X-linked recessive dysmyelinating disorder of the central nervous system in which myelin is not formed properly. It is characterized clinically by nystagmus, spastic quadriplegia, ataxia, and developmental delay.
    Defects in PLP1 are the cause of spastic paraplegia X-linked type 2 (SPG2) [MIM:312920]. SPG2 is characterized by spastic gait and hyperreflexia. In some patients, complicating features include nystagmus, dysarthria, sensory disturbance, mental retardation, optic atrophy.
  • Sequence similarities
    Belongs to the myelin proteolipid protein family.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • HLD1 antibody
    • Lipophilin antibody
    • Major myelin proteolipid protein antibody
    • MMPL antibody
    • Myelin proteolipid protein antibody
    • MYPR_HUMAN antibody
    • PLP 1 antibody
    • PLP antibody
    • PLP/DM20 antibody
    • PLP1 antibody
    • PMD antibody
    • Proteolipid protein 1 antibody
    • SPG2 antibody
    see all

Images

  • ab28486 staining Myelin PLP in Rabbit brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with methanol, permeablized with acetone and blocked with 5% BSA for 1 hour at 37°C. The sample was incubated with primary antibody (1/100) at 4°C for 18 hours. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Immunofluorescence analysis of Human brain cells, staining Myelin PLP with ab28486.

    Cells were fixed with formaldehyde and blocked with 10% serum for 20 minutes at room temperature. Samples were incubated with primary antibody (1/400 in diluent) for 16 hours. An AlexaFluor®555-conjugated donkey anti-rabbit polyclonal IgG (1/1000) was used as the secondary antibody.

    See Abreview

  • Immunohistochemical staining of  Myelin PLP in mouse brain thick section using ab28486 (1/1000 dilution). Staining (m) is limited to myelin tracts and oligodendrocytes.

  • ab28486 at a 1/1000 dilution staining mouse brain (white matter) by IHC-P. The tissue was fixed in 10% formalin saline. Antigen retrieval was via hydrated autoclaving at 121°C for 15 minutes followed by 5 minutes in 98%formic acid. The antibody was incubated with the tissue for 16 hours and then bound antibody was detected using a biotinylated goat anti-rabbit IgG Fc.

    See Abreview

References

This product has been referenced in:
  • Farhangi S  et al. In vivo conversion of astrocytes to oligodendrocyte lineage cells in adult mice demyelinated brains by Sox2. Mult Scler Relat Disord 28:263-272 (2019). Read more (PubMed: 30639828) »
  • Radulescu CI  et al. Manipulation of microbiota reveals altered callosal myelination and white matter plasticity in a model of Huntington disease. Neurobiol Dis 127:65-75 (2019). Read more (PubMed: 30802499) »
See all 27 Publications for this product

Customer reviews and Q&As

11-16 of 16 Abreviews or Q&A

Question

Hello,

Please find the possible details in this reply along with the image of the gels attached as a power point.

Looking forward for an answer.

With regards,

Bandita


Order Details
Antibody code: ab28486

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background (Multiple bands , 10kD and 42 kd bands cannot be explained, 30 and 20kd band is expected to correspond to PLP and DM20 proteins)

Lot number

General Information
Antibody storage conditions (temperature/reconstitution etc) -20C after reconstitution


Description of the problem (high background, wrong band size, more bands, no band etc.) more bands


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Mouse myelin protein


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) 1XLDS buffer, with 8 min of heating @ 90C.


Amount of protein loaded: 5 microgram


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) 4-12% gardient bis-tris gel, run in 1X MOPS buffer. Use of beta-mercaptoethanol in sample buffer and running buffer, shows 43kd, 30kd and 10kd resistant bands. The 20kd band totally disappears.


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.), Tris-Glycine transfer buffer@ low voltage, 4C temperature, 4-5 hrs. Blocking 5% non-fat milk in TBST at room temperature for 2hrs.


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Primary 1:1000 overnight @4C. TBST wash 5x 10min at room temperature


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): anti-rabbit secondary tagged with HRP, TBST 5x10 min wash.


Detection method (ECL, ECLPlus etc.) ECL


Positive and negative controls used (please specify): The first condition described includes the positive control



Optimization attempts (problem solving): none
How many times have you tried the Western?: Twice



Have you run a "No Primary" control? No
Yes No

Do you obtain the same results every time? No there has been variability.
Yes No
e.g. are the background bands always in the same place?


What steps have you altered? Reduced the amount of protein loaded and a more purified form of protein was loaded.


Additional Notes: 4 bands are obatined (42-43, 30, 20 and 10kd). Theoretically this PLP antibody is developed against an epitope which corresponds to both PLP and its alternative splice-isoform DM20. Hence I expect that it will identify the protein bands of ˜ 30 and 20KD, 43 kd band and 10kd is unexplained. If I can assume that 43 kd band is a non-specific band, i could be resonable. But when I use reducing conditions during sample preparation and in the running buffer, 43, 30 and 10kd band persists, whereas th 20kd band disappears.


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More
Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab28486. I would also appreciate if you can confirm some further details:

1. I am sorry to confirm that the order number provided was from nearly 4 years ago. I am sorry this is past the 6 month guarantee period, we are not able to offer any credit or free of charge replacments on this occasion. We would encourage customer to contact us as soon as possible if they have difficulties with the results from a product, and before 6 moths after purchase.

2. I am sorry I am not sure of the sample that has been used. Please confirm:
How has the protein been purfied?
Or, have you run cell lysate? In which case which sample were used (which tissue or cell line from mouse). How was the cell lysate prepared? which lyssi buffer did you use?

3. I can recommend to try fresh samples and fresh buffers, checking the pH.

4. I would suggest it is better to reduce with beta mercaptoethanol.
It may be that the DM20 runs at 10-15 kDa in its non reduced form.

I hope this information is helpful, thank you for your cooperation. Thank you for your understanding of our guarantee policy, and I look forward to hearing from you with the requested details.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and mouse, rat and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if youare also able to provide an image (including molecular weight markers) which would help us to assess the results.

Regarding the testing details of western blot for this antibody, I have contacted the originator to request further information. As soon as I have this I will send it on to you. However, if we are able to provide a protocol, please notethis will be a guideline only and may require further individual optimization.

Thank you for your time and cooperation. We look forward to receiving the completed questionaire.

Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Spinal cord)
Specification
Spinal cord
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Dr. Karine Thibault

Verified customer

Submitted Sep 27 2011

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult mouse brain)
Specification
Adult mouse brain
Fixative
Paraformaldehyde
Permeabilization
Yes - TBST 1x
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 12 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (spinal cord)
Specification
spinal cord
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate 20 min at 100C
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 24 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain, White matter)
Specification
Brain, White matter
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jul 05 2006

11-16 of 16 Abreviews or Q&A

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